The Human Genome Project (HGP) would benefit immensely from sophisticated, powerful robotic workstations capable of processing and analyzing tens of thousands of samples for mapping and sequencing projects. Several first generation robotic workstations are now commercially available. However, their usefulness to the HUman Genome Project is constrained by hardware and software limitations. This project represents the collaboration between an industrial robotics engineering/biotechnology group and an NIH sponsored human genome center to develop hardware and software for second and third generation robotics for genome applications. The goal of the project is to develop a robotic workstation platform, associated software, and peripheral robotic tools and laboratory materials that allow processing of samples at rates which are at least two orders of magnitude larger than presently available with commercial instruments. Three specific tasks have been targeted for automation: 1) preparation of DNA from cosmid and yeast cultures for mapping, sequencing, PCR amplification or probe preparation, 2) assembly of PCR reactions and amplification using high density arrays, and 3) automated preparation of DNA templates for sequencing and assembly of sequencing reactions. This project has already developed several novel and useful materials and instrumentation for the genome project, including: 1) an 864-well polycarbonate microtitre plate that will serve as the basis for the development of biochemical microreactions and advanced robotics hardware, 2) specialized tools for handling the 864- well format plate with the Beckman Biomek robot, 3) a standalone robotics system allowing centrifuge-based separation, and 4) hardware and protocol development to demonstrate high throughput PCR. When completed, this instrumentation will be the basis of laboratory automation systems capable of processing more than 10,000 samples per day. This new application draws heavily on concepts presented in our prior application (1 R01 HG00611-01). However the significant changes in the scope and approach warranted a new application.